KR102404383B1 - Cosmetic composition for skin anti-aging or anti-wrinkle containing Weissella cibaria cell lysates - Google Patents
Cosmetic composition for skin anti-aging or anti-wrinkle containing Weissella cibaria cell lysates Download PDFInfo
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- KR102404383B1 KR102404383B1 KR1020200063699A KR20200063699A KR102404383B1 KR 102404383 B1 KR102404383 B1 KR 102404383B1 KR 1020200063699 A KR1020200063699 A KR 1020200063699A KR 20200063699 A KR20200063699 A KR 20200063699A KR 102404383 B1 KR102404383 B1 KR 102404383B1
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- South Korea
- Prior art keywords
- skin
- lysate
- culture
- weissella
- lotion
- Prior art date
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Abstract
본 발명은 와이셀라 시바리아 세포 용해물을 포함하는 피부 노화 방지 또는 주름 개선용 화장료 조성물에 관한 것으로, 본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 조성물은 세포독성이 없으며, 콜라겐분해효소 활성을 억제시키고, 미세먼지에 의해 증가된 MMPs의 생성 억제 효과 및 감소된 1형콜라겐(COL1A1)의 합성 증진 효과를 나타내므로, 피부 노화 방지 및 주름 개선을 위한 화장품, 건강기능식품 또는 건강식품 조성물로 유용할 수 있다.The present invention relates to a cosmetic composition for preventing skin aging or improving wrinkles comprising a Weissella cibaria cell lysate, and the Weissella cibaria strain of the present invention, a culture thereof, a lysate thereof, an extract or culture thereof Since the composition containing the cell lysate of water has no cytotoxicity, inhibits collagenase activity, and exhibits an effect of inhibiting the production of MMPs increased by fine dust and reducing the synthesis of type 1 collagen (COL1A1), It may be useful as a cosmetic, health functional food or health food composition for skin aging prevention and wrinkle improvement.
Description
본 발명은 와이셀라 시바리아 세포 용해물을 포함하는 피부 노화 방지 또는 주름 개선용 화장료 조성물에 관한 것으로, 보다 상세하게는 와이셀라 시바리아 세포 용해물을 포함하는 피부 노화 방지 또는 주름 개선용 화장료 조성물에 관한 것이다. The present invention relates to a cosmetic composition for anti-aging or wrinkle improvement comprising a Wasella civaria cell lysate, and more particularly, to a cosmetic composition for preventing skin aging or wrinkle improvement comprising a Wasella civaria cell lysate. it's about
인상은 타인에게 보여지는 모습으로 옷차림과 피부, 표정, 화장, 자세 등으로 형성된다. 현대사회에서 사람의 내면을 인상을 통해 간접적으로 크게 평가되기 때문에 인상을 매우 중요하게 여기고 있으며, 외모를 통해 자신을 표현하는 사람들이 증가하고 있다. 이러한 외모의 중요성이 커짐에 따라 화장품에 대한 수요가 크게 증가하고 있다. 특히 화장품들 중에서도 의약품의 성격도 가지는 주름개선 기능성 화장품에 대한 관심과 수요가 꾸준히 증가하고 있다. Impressions are the appearance that others see, and are formed by clothes, skin, facial expressions, makeup, posture, etc. In modern society, impressions are very important because the inside of a person is greatly evaluated indirectly through impressions, and more and more people express themselves through their appearance. As the importance of appearance increases, the demand for cosmetics is increasing significantly. In particular, among cosmetics, interest and demand for wrinkle-improving functional cosmetics that also have the characteristics of pharmaceuticals are steadily increasing.
최근에는 소비자들이 단순히 완성된 화장품의 효능만 관심을 가지는 것뿐만 아니라 화장품에 사용되는 각각 원료들의 효능 및 안정성에 대한 관심도 커지고 있어 인체에 해로운 화학물질이 첨가된 제품을 피하려는 경향이 커지고 있다. 따라서 많은 화장품 연구소들에서는 인체에 무해한 자연 소재의 기능성 원료 개발 연구가 활발하게 이루어지고 있으며 대부분 최근의 화장품들은 안정성이 보장된 소재 또는 유효 성분들 위주로 사용하여 제품이 만들어지고 있다. 특히 많은 기능성 화장품들 중 주름 개선용 기능성 화장품의 원료 개발은 현재까지도 활발하게 진행중이며 새롭고 다양한 기능성 소재들을 사용한 화장품의 제품 출시율도 증가되고 있다. 최근 몇 년에는 미세먼지와 같은 유해 환경물질로부터 피부를 보호하는 효과를 지닌 여러 종류의 화장품들이 출시되고 있으며 이를 위한 소재 및 제형의 연구개발도 활발하게 수행되고 있다. 이들을 모두 종합하여 보았을 때 안정성이 보장된 자연 유래의 소재가 미세먼지 등 유해 환경 물질 등으로부터의 노화방지 효능을 보이는 연구 개발의 필요성이 대두되고 있다. Recently, consumers are not only interested in the efficacy of finished cosmetics, but are also increasingly interested in the efficacy and stability of each raw material used in cosmetics. Therefore, research on the development of functional raw materials of natural materials harmless to the human body is being actively conducted in many cosmetic research institutes, and most recent cosmetics are made using materials with guaranteed stability or active ingredients. In particular, among many functional cosmetics, the development of raw materials for wrinkle-improving functional cosmetics is still in progress, and the launch rate of cosmetics using new and various functional materials is also increasing. In recent years, various types of cosmetics that have the effect of protecting the skin from harmful environmental substances such as fine dust have been released, and research and development of materials and formulations for this purpose are being actively conducted. When all of these are taken together, the need for research and development of natural-derived materials with guaranteed stability to show anti-aging effects from harmful environmental substances such as fine dust is emerging.
현대사회가 급속하게 발전함에 따라 환경오염의 문제도 함께 동반되고 있다. 그 중 대기 오염 물질 중 미세먼지(particulate matter; PM)는 입자가 10 μm이며 질산염(NO3 -), 암모늄(NH4 +), 황산염(SO4 2-) 등 연소 작용에 의해 발생되는 이온성분들과 탄소 화합물, 금속화합물등으로 이루어져 있어 호흡기 질환은 물론 심혈관 질환, 피부 질환 등 각종 여러 질환을 유발하여 인간의 건강에 악영향을 미치고 있어 사회적으로 문제되고 있다. 미세먼지가 제일 먼저 인체에 접촉하는 부위는 피부이다. 미세먼지는 인간의 표피 장벽 기능을 손상시키고 손상된 피부 장벽을 통해 각종 오염물질들이 피부 속으로 침투하게 되어 결과적으로 여드름, 색소침착, 노화 피부염 등을 일으킨다. 더 심하게는 탈모와 피부암을 포함하는 피부질환을 일으킬 수도 있다. With the rapid development of modern society, the problem of environmental pollution is also accompanied. Among air pollutants, particulate matter (PM) has a particle size of 10 μm, and ionic components such as nitrate (NO 3 - ), ammonium (NH 4 + ), and sulfate (SO 4 2- ) are generated by combustion. It is made up of metals, carbon compounds, and metal compounds, causing various diseases, such as respiratory diseases, cardiovascular diseases, and skin diseases, which adversely affect human health, which is a social problem. The first place that fine dust comes into contact with the human body is the skin. Fine dust impairs the human epidermal barrier function, and various contaminants penetrate into the skin through the damaged skin barrier, resulting in acne, pigmentation, and aging dermatitis. Even worse, it can lead to skin diseases including hair loss and skin cancer.
피부 노화를 일으키는 기전으로는 미세먼지, UV 또는 외부 자극에 의해 산화 스트레스가 일어남에 따라 ROS(Reactive oxygen species)가 증가하여 MAPK(Mitogen-activated protein kinase) 기전이 활성화되고 AP-1(Transcription factor activator protein 1) 또는 NF-kB(Nuclear factor-kB) 물질에 영향을 주어 콜라겐을 분해시키는 MMPs(Matrix metalloproteinase)의 발현 및 활성이 증가하여 결과적으로 콜라겐의 합성이 낮아져 피부 노화를 일으킨다고 보고되고 있다. 피부 주름을 형성시키는 요인 중 하나인 미세먼지는 입자가 매우 작아 피부에 쉽게 침투할 수 있어 피부 장벽을 파괴하여 미세먼지 속 오염물질들이 피부면역체계에 영향을 주게 되고 피부 탄력을 유지시키는 콜라겐과 엘라스틴을 파괴시켜 결과적으로 피부의 주름 및 손상을 일으킨다. 이러한 미세먼지를 포함한 외부요인에 의해 피부 상피세포와 진피세포에서 발현 및 활성이 증가된 MMPs는 구조와 기능적 특징에 따라서 크게 Collagenases, Gelatinases, Stromelysins 등으로 구분할 수 있다. Collagenases 에는 MMP-1, -8, -13, -18 이며 이들은 콜라겐구조를 절단하는 역할을 가진다. Glatinases에는 MMP-2와 MMP-9이 존재하며 기저막 콜라겐(Basement membrane collagen)의 구조를 변형시키고 분해한다. MMP-3, -10, -11, -19는 Stromelysins으로써 Proteoglycans, Martix glycoproteins, 콜라겐들을 분해시키고 다양한 MMPs를 활성화시켜 주름 형성에 중요한 역할을 한다. 이러한 미세먼지 의해 증가된 MMPs은 상피세포인 인간 각질형성세포 (Keratinocyte)와 진피세포인 인간 진피섬유아세포 (Human dermal fibroblast) 등에서 발현되어 세포 밖으로 분비되어 결과적으로 진피세포의 콜라겐을 구조적으로 분해시켜 피부탄력이 감소되어 피부 주름을 형성시킨다. 피부 진피층에는 1형콜라겐(COL1A1)이 80~90%를 차지하고 있으며 이 외에 다른 타입의 콜라겐들과 피브로넥틴(fibronectin) 등으로 구성되어 있다. 다양한 외부요인 및 나이가 증가의 이유로 콜라겐분해효소의 활성이 증가하게 되어 콜라겐들을 분해시킴으로써 피부의 매끄러움, 수분, 탄력이 감소된다. 이러한 이유로 콜라겐분해효소의 활성 및 콜라겐 유전자의 감소와 MMPs의 발현 증가는 주름 생성에 주요 요인으로 콜라겐분해효소의 활성 억제 및 콜라겐과 MMPs 유전자들의 발현 변화를 통해 물질의 주름생성완화 효능을 알 수 있다.As the mechanism that causes skin aging, ROS (Reactive oxygen species) increases as oxidative stress occurs by fine dust, UV or external stimuli, the MAPK (Mitogen-activated protein kinase) mechanism is activated, and AP-1 (Transcription factor activator) It has been reported that the expression and activity of matrix metalloproteinase (MMPs) that degrade collagen by affecting protein 1) or NF-kB (nuclear factor-kB) substances increase, resulting in lowered collagen synthesis, resulting in skin aging. Fine dust, one of the factors that causes skin wrinkles, is very small and can easily penetrate the skin, destroying the skin barrier, pollutants in the fine dust affect the skin immune system, and collagen and elastin to maintain skin elasticity It destroys the skin, resulting in wrinkles and damage to the skin. MMPs, whose expression and activity are increased in skin epithelial and dermal cells by external factors including fine dust, can be largely classified into Collagenases, Gelatinases, and Stromelysins according to their structure and functional characteristics. Collagenases include MMP-1, -8, -13, and -18, which have a role in cutting collagen structures. Glatinases include MMP-2 and MMP-9, which modify and degrade the structure of basement membrane collagen. MMP-3, -10, -11, and -19 are Stromelysins, which degrade Proteoglycans, Martix glycoproteins, and collagen, and activate various MMPs to play an important role in wrinkle formation. MMPs increased by such fine dust are expressed in epithelial cells such as human keratinocytes and dermal cells, human dermal fibroblasts, and are secreted out of the cells. The elasticity is reduced and the skin wrinkles are formed. In the dermal layer of the skin,
유산균(Lactic acid bacteria)은 요구르트, 치즈 및 발효유 등 각종 발효 식품 제조에 있어 필수로 사용되고 있는 중요한 미생물이다. 유산균은 식품의 발효뿐만 아니라 사람의 면역력 증진, 식중독 방지, 변비 예방 및 정장효능이 있어 오래전부터 많은 연구개발로 발전을 거듭하였다. 최근에는 암 예방에도 효과가 있다고 보고되고 있다. 이러한 유산균의 엄청난 효능 때문에 다양한 유산균 발효 제품과 음식의 관심이 해를 거듭할수록 높아지고 있으며 그 중 김치는 우리나라의 대표 발효 음식으로 김치에 대한 연구도 끊임없이 수행되고 있다. 김치의 미생물이 완벽하게 밝혀져 있지는 않으나 김치 안에서 발견되는 미생물들은 크게 락토바실러스(Lactobacillus), 와이셀라(Weissella), 류코노스톡(Leuconostoc) 이렇게 크게 세 가지 속(Genus)의 유산균으로 구별된다. 이 중 와이셀라속의 대표 미생물인 와이셀라 시바리아(Weissella cibaria)는 그람양성균으로써 비운동성 단 간균(Short rod)이고, Catalase 음성에다가 호기 또는 혐기 모든 조건에서 잘 자라는 통성혐기성균으로 젖산발효를 하는 큰 특징을 가진다. 또한 와이셀라는 갓김치에 풍부하다고 알려져 있으며 항암, 항 바이러스, 항 비만, 면역 증진, 아토피 개선 등의 효능이 있다고 밝혀져 있지만 화장품 소재로써의 연구는 아직까지 미미하다. Lactic acid bacteria are important microorganisms that are essential for the manufacture of various fermented foods such as yogurt, cheese and fermented milk. Lactic acid bacteria have been developed through research and development for a long time as they not only ferment food, but also enhance human immunity, prevent food poisoning, prevent constipation, and have intestinal effects. Recently, it has been reported to be effective in preventing cancer. Because of the tremendous efficacy of these lactic acid bacteria, interest in various fermented lactic acid bacteria products and foods is increasing year by year. Although the microorganisms of kimchi are not fully known, the microorganisms found in kimchi are largely divided into three genera: Lactobacillus, Weissella , and Leuconostoc . Among them, Weissella cibaria , a representative microorganism of the genus Weissella, is a Gram-positive, non-motile short rod, Catalase-negative, and a facultative anaerobic bacterium that grows well in both aerobic and anaerobic conditions. have a characteristic In addition, Wasella is known to be abundant in fresh kimchi, and has been found to have anticancer, antiviral, anti-obesity, immune enhancement, and atopy improvement effects, but research as a cosmetic material is still insignificant.
이에 본 발명자들은 와이셀라 시바리아 세포 용해물이 콜라겐분해효소의 활성을 억제시키고 미세먼지로 인한 주름생성관련 유전자들의 발현을 조절함으로써 미세먼지로 인한 피부 노화 방지 및 피부 주름 개선 효과를 확인하고 본 발명을 완성하였다.Therefore, the present inventors confirmed the effect of preventing skin aging and improving skin wrinkles due to fine dust by inhibiting the activity of the Weissella civaria cell lysate and regulating the expression of genes related to wrinkle generation due to fine dust, and was completed.
본 발명의 목적은 피부 주름 개선 또는 탄력증진용 화장료 조성물을 제공하는 것이다.It is an object of the present invention to provide a cosmetic composition for improving skin wrinkles or enhancing elasticity.
본 발명의 다른 목적은 피부 노화 예방 또는 개선용 화장료 조성물을 제공하는 것이다.Another object of the present invention is to provide a cosmetic composition for preventing or improving skin aging.
본 발명의 또 다른 목적은 피부 주름 개선 또는 탄력증진용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for improving skin wrinkles or enhancing elasticity.
본 발명의 다른 목적은 피부 노화 예방 또는 개선용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for preventing or improving skin aging.
본 발명의 또 다른 목적은 피부 주름 개선 또는 탄력증진용 건강식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health food composition for improving skin wrinkles or enhancing elasticity.
본 발명의 다른 목적은 피부 노화 예방 또는 개선용 건강식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health food composition for preventing or improving skin aging.
상기 목적을 달성하기 위하여,In order to achieve the above object,
본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 주름 개선 또는 탄력증진용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin wrinkles or enhancing elasticity, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
또한, 본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 노화 예방 또는 개선용 화장료 조성물을 제공한다.In addition, the present invention provides a cosmetic composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
나아가 본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 주름 개선 또는 탄력증진용 건강기능식품 조성물을 제공한다.Furthermore, the present invention provides a health functional food composition for improving skin wrinkles or enhancing elasticity, including a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
더 나아가 본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 노화 예방 또는 개선용 건강기능식품 조성물을 제공한다.Furthermore, the present invention provides a health functional food composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
또한, 본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 주름 개선 또는 탄력증진용 건강식품 조성물을 제공한다.In addition, the present invention provides a health food composition for improving skin wrinkles or enhancing elasticity, including a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
나아가 본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 노화 예방 또는 개선용 건강식품 조성물을 제공한다.Furthermore, the present invention provides a health food composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 조성물은 세포독성이 없으며, 콜라겐분해효소 활성을 억제시키고, 미세먼지에 의해 증가된 MMPs의 생성 억제 효과 및 감소된 1형콜라겐(COL1A1)의 합성 증진 효과를 나타내므로, 피부 노화 방지 및 주름 개선을 위한 화장품, 건강기능식품 또는 건강식품 조성물로 유용할 수 있다.The composition comprising the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or cell lysate of the culture is not cytotoxic, and inhibits collagenase activity, Since it exhibits an increased inhibitory effect on the production of MMPs and a reduced effect on the synthesis of collagen type 1 (COL1A1), it may be useful as a cosmetic, health functional food or health food composition for skin aging prevention and wrinkle improvement.
도 1은 본 발명의 와이셀라 세포 용해물을 인간 각질형성세포(HaCaT), 인간 섬유아세포(CCD-986Sk), 인간 진피 섬유아세포(HDF-1)에서 세포독성을 확인한 결과이다.
도 2는 본 발명의 와이셀라 세포 용해물의 콜라겐분해효소의 활성 억제 효능을 확인한 결과이다.
도 3은 인간 각질형성세포(HaCaT)에서 본 발명의 와이셀라 세포 용해물의 미세먼지에 의해 증가된 MMPs 발현 억제 효능을 확인한 결과이다.
도 4는 인간 섬유아세포(CCD-986Sk) 및 인간 진피 섬유아세포(HDF-1)에서 본 발명의 와이셀라 세포 용해물의 미세먼지에 의해 감소된 1형콜라겐(COL1A1) 발현 증가 효능을 확인한 결과이다.1 is a result of confirming the cytotoxicity of the Weissella cell lysate of the present invention in human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1).
Figure 2 is the result of confirming the activity inhibitory effect of the collagen degrading enzyme of the Weissella cell lysate of the present invention.
3 is a result confirming the inhibitory efficacy of MMPs expression increased by fine dust in the Weissella cell lysate of the present invention in human keratinocytes (HaCaT).
4 is a result confirming the effect of increasing the expression of collagen type 1 (COL1A1) reduced by fine dust in the Weissella cell lysate of the present invention in human fibroblasts (CCD-986Sk) and human dermal fibroblasts (HDF-1) .
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
피부 주름 개선 또는 탄력증진용 화장료 조성물Cosmetic composition for improving skin wrinkles or enhancing elasticity
본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 주름 개선 또는 탄력증진용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin wrinkles or enhancing elasticity, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
본 발명의 일실시예에 있어서, 상기 조성물은 균주, 이의 배양물, 이의 파쇄물, 이의 추출물을 포함하거나 배양물의 세포 현탁액, 세포 균질물, 세포 용해물, 세포 상청액, 세포 여액, 세포 펠렛 또는 하나 이상의 세포들이거나, 또는 이를 포함하는 것일 수 있다. 보다 바람직하게 와이셀라 시바리아(Weissella cibaria) 균주 배양물의 세포 용해물일 수 있다.In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.
본 발명의 화장료 조성물에 있어서, 상기 피부 주름은 미세먼지에 의한 것일 수 있다. In the cosmetic composition of the present invention, the skin wrinkles may be caused by fine dust.
노화에는 콜라겐 분해효소인 기질 금속 단백질 분해 효소(Matrix Metalloproteinase; MMP)라는 효소도 관여한다. 즉, 생체 내에서 콜라겐과 같은 세포외 기질의 합성과 분해는 적절하게 조절되고 있는데, 노화가 진행되면서 콜라겐 합성이 감소하고, 콜라겐을 분해하는 효소인 기질 금속 단백질 분해 효소(MMP)의 발현이 촉진되어 피부의 탄력이 저하되고 주름이 형성된다. Aging also involves an enzyme called Matrix Metalloproteinase (MMP), a collagen degrading enzyme. That is, the synthesis and decomposition of extracellular matrix such as collagen in vivo is properly regulated. As aging progresses, collagen synthesis decreases, and expression of matrix metalloproteinase (MMP), an enzyme that decomposes collagen, is promoted. As a result, the elasticity of the skin decreases and wrinkles are formed.
그러므로 세포 내에서 활성화가 유도되는 MMP 발현을 조절하거나 그 활성을 억제할 수 있는 물질의 경우, 피부 탄력 개선, 주름개선 및 노화 억제를 위한 기능성 화장료의 소재로 활용할 수 있다. Therefore, in the case of a substance capable of regulating or inhibiting MMP expression induced in activation in cells, it can be used as a material for functional cosmetics for improving skin elasticity, improving wrinkles, and inhibiting aging.
콜라겐 분해를 촉매하는 기질금속단백질 분해효소(matrixmetalloproteinase, MMP)는 포유류에서 23개의 타입이 존재하며 이중 자외선에 의해 증가하는 MMP 타입은 1, 3 및 9번으로 알려져 있고 이들 세가지 타입의 MMPs는 콜라겐 타입 1과 3을 분해하는 효소이다. 구체적으로, MMP-1의 경우, 콜라겐섬유의 중간을 절단하며 MMP-3, MMP-9는 상기 절단된 콜라겐섬유를 세분해서 절단하는 역할을 한다. There are 23 types of matrix metalloproteinase (MMP) that catalyze collagen degradation in mammals. Among them, MMP types increased by UV light are known as 1, 3, and 9, and these three types of MMPs are collagen type. It is an enzyme that breaks down 1 and 3. Specifically, in the case of MMP-1, the middle of the collagen fiber is cut, and MMP-3 and MMP-9 serve to subdivide and cut the cut collagen fiber.
즉, 본 발명에 따른 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 조성물은 MMPs 유전자의 발현을 유의적으로 감소시킴으로써, 콜라겐 섬유의 분해를 억제할 수 있으며, 피부조직에서 콜라겐의 합성을 증가시킴에 따라, 피부 탄력 개선, 주름개선 및 노화를 억제할 수 있다(실험예 3 참조). That is, the composition comprising the Weissella cibaria strain, its culture, its lysate, its extract, or cell lysate of the culture according to the present invention significantly reduces the expression of the MMPs gene, so that the Decomposition can be inhibited, and by increasing the synthesis of collagen in the skin tissue, skin elasticity improvement, wrinkle improvement, and aging can be suppressed (see Experimental Example 3).
본 발명에 따른 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클린저로 구성된 군으로부터 선택되는 1종 이상의 제형인 것일 수 있고, 피부 외용제 형태로도 사용할 수 있다. 상기한 필수 성분 이외의 다른 성분들은 기타 제형 또는 사용 목적 등에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있다.The cosmetic composition according to the present invention includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, It may be one or more formulations selected from the group consisting of soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser, and may also be used in the form of external preparations for skin. Components other than the above essential components can be appropriately selected and formulated by those skilled in the art without difficulty depending on other formulations or purposes of use.
본 발명의 화장료 조성물은 바람직하게는 화장품학적으로 허용가능한 매질 또는 기제를 함유한다. 이는 국소적용에 적합한 모든 제형으로, 예를 들면 용액, 겔, 고체 또는 반죽 무수 생성물, 수상에 유상을 분산시켜 얻은 에멀젼, 현탁액 마이크로 에멀젼, 마이크로캡슐, 미세과립구 또는 이온형(리포좀) 및/또는 바이온형의 소낭 분산제의 형태로, 또는 크림, 스킨, 로션, 파우더, 연고, 스프레이 또는 콘실 스틱의 형태로 제공될 수 있다. 또한, 포말(foam)의 형태 또는 압축된 추진제를 더 함유한 에어로졸 조성물의 형태로도 제조될 수 있다. 또한, 본 발명의 화장료 조성물은 피부과학적으로 허용가능한 매질 또는 기제를 함유함으로써 피부과학 분야에서 통상적으로 사용되는 국소적용 또는 전신적용할 수 있는 보조제 형태로 제조될 수 있으며, 이들 조성물은 당해 분야의 통상적 방법에 따라 제조될 수 있다.The cosmetic composition of the present invention preferably contains a cosmetically acceptable medium or base. It is in any dosage form suitable for topical application, for example solutions, gels, solid or kneaded dry products, emulsions obtained by dispersing the oily phase in an aqueous phase, suspension microemulsions, microcapsules, microgranules or ionic (liposomes) and/or bi It may be provided in the form of a warm, vesicular dispersion, or in the form of a cream, toner, lotion, powder, ointment, spray or cone stick. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant. In addition, the cosmetic composition of the present invention can be prepared in the form of an adjuvant that can be applied topically or systemically commonly used in the field of dermatology by containing a dermatologically acceptable medium or base, and these compositions are conventional in the art. It can be prepared according to the method.
또한, 상기 화장료 조성물은 본 발명의 추출물에 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제(foaming agent), 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 또는 화장품에 통상적으로 사용되는 임의의 다른 성분과 같은 화장품 분야에서 통상적으로 사용되는 보조제를 함유할 수 있다. 이들 보조제는 화장품학 분야에서 일반적으로 사용되는 양으로 도입된다.In addition, the cosmetic composition includes a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, and water in addition to the extract of the present invention. , ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or commonly used in cosmetics It may contain adjuvants commonly used in the cosmetic field, such as any other ingredients. These adjuvants are introduced in amounts generally used in the field of cosmetology.
피부 노화 예방 또는 개선용 화장료 조성물Cosmetic composition for preventing or improving skin aging
본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 노화 예방 또는 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
본 발명의 일실시예에 있어서, 상기 조성물은 균주, 이의 배양물, 이의 파쇄물, 이의 추출물을 포함하거나 배양물의 세포 현탁액, 세포 균질물, 세포 용해물, 세포 상청액, 세포 여액, 세포 펠렛 또는 하나 이상의 세포들이거나, 또는 이를 포함하는 것일 수 있다. 보다 바람직하게 와이셀라 시바리아(Weissella cibaria) 균주 배양물의 세포 용해물일 수 있다.In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.
본 발명의 화장료 조성물에 있어서, 상기 피부 주름은 미세먼지에 의한 것일 수 있다. In the cosmetic composition of the present invention, the skin wrinkles may be caused by fine dust.
본 발명에 따른 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클린저로 구성된 군으로부터 선택되는 1종 이상의 제형인 것일 수 있고, 피부 외용제 형태로도 사용할 수 있다. 상기한 필수 성분 이외의 다른 성분들은 기타 제형 또는 사용 목적 등에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있다.The cosmetic composition according to the present invention includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, It may be one or more formulations selected from the group consisting of soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser, and may also be used in the form of external preparations for skin. Components other than the above essential components can be appropriately selected and formulated by those skilled in the art without difficulty depending on other formulations or purposes of use.
피부 주름 개선 또는 탄력증진용 건강기능식품 또는 건강식품 조성물Health functional food or health food composition for skin wrinkle improvement or elasticity enhancement
본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 주름 개선 또는 탄력증진용 건강기능식품 또는 건강식품 조성물을 제공한다.The present invention provides a health functional food or health food composition for improving skin wrinkles or enhancing elasticity, including a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.
본 발명의 일실시예에 있어서, 상기 조성물은 균주, 이의 배양물, 이의 파쇄물, 이의 추출물을 포함하거나 배양물의 세포 현탁액, 세포 균질물, 세포 용해물, 세포 상청액, 세포 여액, 세포 펠렛 또는 하나 이상의 세포들이거나, 또는 이를 포함하는 것일 수 있다. 보다 바람직하게 와이셀라 시바리아(Weissella cibaria) 균주 배양물의 세포 용해물일 수 있다. In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.
본 발명의 조성물에 있어서, 상기 피부 주름은 미세먼지에 의한 것일 수 있다. In the composition of the present invention, the skin wrinkles may be caused by fine dust.
본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 건강기능식품 및 건강식품 조성물로 사용하는 경우, 식품의 종류에는 특별한 제한은 없다. 본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 첨가할 수 있는 건강기능식품의 예로는 정제, 캡슐제, 환제 또는 액제 형태일 수 있으며, 건강식품의 예로는 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강기능식품 및 건강식품 조성물을 모두 포함한다.When the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or cell lysate of the culture is used as a health functional food and health food composition, the type of food is not particularly limited. Examples of health functional food to which the Weissella cibaria strain of the present invention, its culture, its lysate, its extract, or cell lysate of the culture can be added may be in the form of tablets, capsules, pills or liquids. Examples of health food include various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic beverages, beverages, alcoholic beverages, gum, tea and vitamin complexes, etc. of health functional food and health food composition.
본 발명에 따른 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 함유하는 건강기능식품 및 건강식품 조성물은 식품에 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물의 혼합량은 그의 사용 목적(예방 또는 개선용)에 따라 적합하게 결정될 수 있다. 일반적으로, 건강기능식품 및 건강식품 조성물 중의 상기 조성물의 양은 전체 식품 중량의 0.1 내지 90 중량부로 가할 수 있다. 그러나 건강 유지를 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물은 상기 범위 이상의 양으로도 사용될 수 있다. Weissella cibaria strain according to the present invention, its culture, its lysate, its extract or health functional food and health food composition containing the cell lysate of the culture is added to food or other food or food ingredients can be used together, and can be appropriately used according to a conventional method. Weissella cibaria strain, its culture, its lysate, its extract, or the mixed amount of the cell lysate of the culture may be suitably determined depending on the purpose of its use (for prevention or improvement). In general, the amount of the composition in the health functional food and health food composition may be added in an amount of 0.1 to 90 parts by weight based on the total weight of the food. However, in the case of long-term intake for health maintenance or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the Weissella cibaria strain, its culture, A lysate thereof, an extract thereof, or a cell lysate of a culture may be used in an amount greater than or equal to the above range.
본 발명의 건강기능식품 및 건강식품 조성물은 지시된 비율로 필수 성분으로서 본 발명 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 함유하는 것 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트라이톨 등의 당알코올이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 건강기능식품 및 건강식품 조성물 100 당 일반적으로 약 1 내지 20 g, 바람직하게는 약 5 내지 12 g이다.The health functional food and health food composition of the present invention is an essential ingredient in the indicated ratio, and the present invention Weissella cibaria strain, its culture, its lysate, its extract or cell lysate of the culture. The other ingredients are not particularly limited and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 health functional food and health food composition of the present invention.
상기 외에 본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 함유하는 건강기능식품 및 건강식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강기능식품 및 건강식품 조성물은 천연 과일쥬스 및 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다.In addition to the above, the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or health functional food and health food composition containing a cell lysate of the culture are various nutrients, vitamins, minerals (electrolytes) ), synthetic and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives , glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like. In addition, the health functional food and health food composition of the present invention may contain natural fruit juice, fruit juice for the production of fruit juice drinks, and vegetable drinks.
이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 함유하는 건강기능식품 및 건강식품 조성물 100 중량부 당 0.1 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.These components may be used independently or in combination. Although the ratio of these additives is not so important, the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or 100 parts by weight of a health food composition containing a cell lysate of the extract or culture It is generally selected in the range of 0.1 to about 20 parts by weight of sugar.
피부 노화 예방 또는 개선용 건강기능식품 또는 건강식품 조성물Health functional food or health food composition for preventing or improving skin aging
본 발명은 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 포함하는 피부 노화 예방 또는 개선용 건강기능식품 또는 건강식품 조성물을 제공한다.The present invention provides a health functional food or health food composition for preventing or improving skin aging, comprising a Weissella cibaria strain, its culture, its lysate, its extract, or a cell lysate of the culture.
본 발명의 일실시예에 있어서, 상기 조성물은 균주, 이의 배양물, 이의 파쇄물, 이의 추출물을 포함하거나 배양물의 세포 현탁액, 세포 균질물, 세포 용해물, 세포 상청액, 세포 여액, 세포 펠렛 또는 하나 이상의 세포들이거나, 또는 이를 포함하는 것일 수 있다. 보다 바람직하게 와이셀라 시바리아(Weissella cibaria) 균주 배양물의 세포 용해물일 수 있다.In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.
본 발명의 조성물에 있어서, 상기 피부 주름은 미세먼지에 의한 것일 수 있다. In the composition of the present invention, the skin wrinkles may be caused by fine dust.
본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 건강기능식품 및 건강식품 조성물로 사용하는 경우, 식품의 종류에는 특별한 제한은 없다. 본 발명의 와이셀라 시바리아(Weissella cibaria) 균주, 이의 배양물, 이의 파쇄물, 이의 추출물 또는 배양물의 세포 용해물을 첨가할 수 있는 건강기능식품의 예로는 정제, 캡슐제, 환제 또는 액제 형태일 수 있으며, 건강식품의 예로는 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강기능식품 및 건강식품 조성물을 모두 포함한다.When the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or cell lysate of the culture is used as a health functional food and health food composition, there is no particular limitation on the type of food. Examples of health functional foods to which the Weissella cibaria strain of the present invention, its culture, its lysate, its extract, or cell lysate of the culture can be added may be in the form of tablets, capsules, pills or liquids. Examples of health food include various drinks, meat, sausages, bread, candies, snacks, noodles, ice cream, dairy products, soups, ionic beverages, beverages, alcoholic beverages, gum, tea and vitamin complexes, etc. of health functional food and health food composition.
하기의 실시예를 통하여 본 발명을 보다 상세하게 설명한다. 그러나 하기 실시예는 본 발명의 내용을 구체화하기 위한 것일 뿐 이에 의해 본 발명이 한정되는 것은 아니다.The present invention will be described in more detail through the following examples. However, the following examples are only intended to embody the contents of the present invention, and the present invention is not limited thereto.
<실시예 1> 와이셀라 시바리아 세포 용해물 준비<Example 1> Preparation of Weissella civaria cell lysate
본 발명에 사용된 와이셀라 시바리아는 시중에 판매하는 김치로부터 분리하였다. 김치를 잘게 파쇄하고 여과하여 MRS agar (Lactobacilli MRS agar)배지에 도말 후 16시간 배양하였다. 배양된 여러 종류의 콜로니를 선별 후 MRS borth에 추가 배양하고 16S ribosoma RNA sequence 분석을 수행하였다. 분석된 균주들 중 피부 노화 및 주름개선 효능이 아직 밝혀져 있지않은 와이셀라 시바리아 균을 선택하여 MRS borth 배지에 접종하고 30℃에서 24시간 배양하여 본 연구에 사용하였다.Weissella civaria used in the present invention was isolated from commercially available kimchi. The kimchi was crushed finely, filtered, spread on MRS agar (Lactobacilli MRS agar), and cultured for 16 hours. After selecting several types of cultured colonies, they were further cultured in the MRS borth, and 16S ribosoma RNA sequence analysis was performed. Among the analyzed strains, Weissella civaria bacteria, whose efficacy in improving skin aging and wrinkle have not yet been identified, were selected and inoculated in MRS borth medium and cultured at 30° C. for 24 hours for use in this study.
와이셀라 시바리아를 MRS agar 배지(Lactobacilli MRS agar)에 도말 후 30℃에서 16시간 배양하여 단일 콜로니를 얻었다. 얻은 단일 콜로니를 MRS 액체배지(Lactobacilli MRS broth)에 접종한 후 30℃에서 16시간 배양 후, 균체를 7,000 rpm에서 10분 동안 원심분리하여 배양액을 제거하고, PBS(Phosphate-buffered Saline) 버퍼로 3회 세척하여 배지 성분을 제거하였다. 배지 성분이 제거된 세포만 원심분리 하여 얻은 후 1.5 ㎖ 튜브에 분주 후 PBS 버퍼 400㎕를 첨가 하여 소니케이터로 세포를 용해시키고 13,000 rpm에서 5분간 원심분리 하여 세포용해물인 상등액만 새로운 튜브에 옮겨 단백질 정량 후 사용하였다. 본 발명에 사용된 와이셀라 시바리아 세포 용해물을 이하 와이셀라 용해물이라 칭하였다.After spreading Weissella civaria on MRS agar medium (Lactobacilli MRS agar), it was cultured at 30° C. for 16 hours to obtain a single colony. After the obtained single colony was inoculated in Lactobacilli MRS broth, incubated at 30° C. for 16 hours, the cells were centrifuged at 7,000 rpm for 10 minutes to remove the culture medium, and PBS (Phosphate-buffered Saline) buffer was used for 3 Washed times to remove media components. Only the cells from which the medium component has been removed are obtained by centrifugation, and after dispensing into 1.5 ml tubes, 400 μl of PBS buffer is added to lyse the cells with a sonicator, and centrifuged at 13,000 rpm for 5 minutes. It was transferred and used after protein quantification. The Weissella civaria cell lysate used in the present invention was hereinafter referred to as the Weissella lysate.
<준비예 1> 세포배양<Preparation Example 1> Cell culture
실험에 사용된 인간피부세포인 인간 각질형성세포(HaCaT), 인간 섬유아세포(CCD-986Sk), 인간 진피 섬유아세포(HDF-1)들은 37℃, CO2 인큐베이터에서 배양하였으며 3일에 한번씩 새로운 DMEM(Dulbecco Modified Eagle Medium)으로 교체하거나 PBS로 세척 후 Tyrpsin-EDTA 용액을 넣어 세포를 배양접시로부터 분리 후 계대배양 하여 유지하였다.Human skin cells used in the experiment, such as human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1), were cultured in an incubator at 37 ° C., CO 2 and fresh DMEM once every 3 days. (Dulbecco Modified Eagle Medium) or after washing with PBS, Tyrpsin-EDTA solution was added to separate the cells from the culture dish and subcultured and maintained.
<실험예 1> 세포독성 확인<Experimental Example 1> Confirmation of cytotoxicity
인간 피부세포에서 와이셀라 시바리아 세포 용해물의 독성을 확인하기 위하여, 인간 각질형성세포(HaCaT), 인간 섬유아세포(CCD-986Sk), 인간 진피 섬유아세포(HDF-1)들을 각 24웰-플레이트에 같은 세포량으로 분주하여 37℃ CO2 인큐베이터에서 24시간 동안 배양하였다. 배양 후 와이셀라 용해물을 농도별로 (0 ㎍/㎖, 100 ㎍/㎖, 200 ㎍/㎖) 처리한 후 37℃ CO2 인큐베이터에서 24시간 동안 추가 배양하였다. 배양이 끝난 후 PBS 용액으로 세척 후 Trypsin-EDTA 용액을 100 ㎕ 씩 첨가 후 같은 조건의 인큐베이터에서 3분간 배양하여 세포를 완전히 배양접시로부터 떨어트린 후 DMEM 배지를 각 각 400 ㎕씩 첨가 후 파이펫을 이용하여 세포를 잘 섞어 주었다. 세포 수를 세기 위해 세포 10 ㎕를 Hemocytometer에 로딩하여 살아있는 세포 수를 계산하였다.To determine the toxicity of Weissella cibaria cell lysates in human skin cells, human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1) were each in a 24-well-plate. Incubated for 24 hours in a 37 ℃ CO 2 incubator by dividing the same amount of cells. After culturing, the Wasella lysate was treated by concentration (0 μg/ml, 100 μg/ml, 200 μg/ml), and then further cultured in a 37° C. CO 2 incubator for 24 hours. After incubation, wash with PBS solution, add 100 μl of Trypsin-EDTA solution, incubate for 3 minutes in the incubator under the same conditions, completely remove the cells from the culture dish, add 400 μl of DMEM medium each, and pipette cells were mixed well. To count the cells, 10 μl of cells were loaded into a hemocytometer and the number of viable cells was counted.
인간 각질형성세포(HaCaT), 인간 섬유아세포(CCD-986Sk), 인간 진피 섬유아세포(HDF-1)에서 와이셀라 용해물의 세포독성을 확인한 결과, 상기 표 1 및 도 1에 나타낸 바촤 같이, 100 ㎍/㎖ 농도에서 독성을 나타내지 않았다. 또한, 200 ㎍/㎖의 농도로 와이셀라 용해물 처리시에는 HaCaT 세포에서 독성을 보인 반면, 다른 두 세포들에서는 크게 독성을 보이지 않았다. 이후 실험에서 와이셀라 용해물을 100 ㎍/㎖의 농도로 사용하였다. As a result of confirming the cytotoxicity of Weissella lysates in human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1), as shown in Table 1 and Figure 1, 100 There was no toxicity at the μg/ml concentration. In addition, when the Weissella lysate was treated at a concentration of 200 μg/ml, it was toxic to HaCaT cells, but not significantly toxic to the other two cells. In subsequent experiments, the Weissella lysate was used at a concentration of 100 μg/ml.
<실험예 2> 와이셀라 시바리아 세포 용해물의 콜라겐분해효소 활성 억제 확인<Experimental Example 2> Confirmation of inhibition of collagenase activity of Wasella civaria cell lysate
와이셀라 용해물이 콜라겐을 분해하는 콜라겐분해효소 활성을 억제시키는지 확인하기위해 96-웰 플레이트에 다양한 농도(0, 40, 80 및 160 ㎍/㎖)의 와이셀라 용해물과 기질인 0.6mM FALGPA(N-[3-(2-Furyl)acryloyl]-Leu-Gly-Pro-Ala)를 400mM NaCl2 와 10mM CaCl2가 포함된 50mM Tricine buffer (pH 7.5)에서 15분간 상온에서 반응시킨 후 0.2 units/ml 콜라겐분해효소(Collagenase)를 각각 넣고 345nm 파장에서 매분마다 흡광도를 측정하여 기질이 콜라겐분해효소에 의해 분해되는 정도를 측정하였다. 여기서 음성대조군은 기질과 버퍼만 넣은 블랭크를 의미한다.To determine whether the Wasella lysate inhibits collagenase activity that degrades collagen, various concentrations (0, 40, 80, and 160 μg/ml) of the Wasella lysate and the substrate 0.6 mM FALGPA were placed in 96-well plates. (N-[3-(2-Furyl)acryloyl]-Leu-Gly-Pro-Ala) was reacted in 50mM Tricine buffer (pH 7.5) containing 400mM NaCl 2 and 10mM CaCl 2 at room temperature for 15 minutes and then 0.2 units /ml Collagenase (Collagenase) was added, respectively, and absorbance was measured every minute at a wavelength of 345 nm to measure the degree of decomposition of the substrate by the collagenase. Here, the negative control group refers to a blank containing only the substrate and the buffer.
그 결과, 도 2에 나타낸 바와 같이, 와이셀라 용해물은 농도 의존적으로 콜라겐 분해효소의 활성을 감소시키는 것을 확인하였다. 상기 실험 결과를 통해 본 발명의 와이셀라 용해물이 콜라겐분해효소의 활성을 억제시킴으로써 피부 노화 방지 및 피부 주름 개선 효과를 갖는 것을 알 수 있었다.As a result, as shown in FIG. 2 , it was confirmed that the Weissella lysate decreased the activity of the collagenase in a concentration-dependent manner. Through the above experimental results, it was found that the Wasella lysate of the present invention had the effect of preventing skin aging and improving skin wrinkles by inhibiting the activity of collagen degrading enzyme.
<실험예 3> 와이셀라 시바리아 세포 용해물의 미세먼지에 의한 MMPs 및 콜라겐 유전자의 발현 확인<Experimental Example 3> Confirmation of expression of MMPs and collagen genes by fine dust of Weissella civaria cell lysate
와이셀라 용해물이 인간 피부 세포에서 피부 노화에 관여하는 유전자들의 발현을 변화시키는지 확인하기 위해 세포들을 각 각 6웰-플레이트에 동일 세포수로 분주하여 37℃ CO2 인큐베이터에서 24시간 동안 배양하였다. 배양 후 와이셀라 용해물을 농도별(0, 50 및 100㎍/㎖)로 처리한 후 37℃ CO2 인큐베이터에서 1시간 동안 전처리 후 미세먼지를 첨가하여 16시간 동안 추가 배양하였다. 본 실험에서 미세먼지는 NIST(National Institute of Standards and Technology)의 SRM(Standard Reference Material) 표준물질인 4 μm 이하의 미세먼지(NIST2786)를 처리하였다. In order to determine whether the Weissella lysate changes the expression of genes involved in skin aging in human skin cells, the cells were each divided into 6 well-plates with the same number of cells and cultured in a 37° C. CO 2 incubator for 24 hours. . After culturing, the Wasella lysate was treated at different concentrations (0, 50 and 100 μg/ml), and then pre-treated in a 37° C. CO 2 incubator for 1 hour, fine dust was added, and further cultured for 16 hours. In this experiment, fine dust of 4 μm or less (NIST2786), which is a standard reference material (SRM) of the National Institute of Standards and Technology (NIST), was treated for fine dust.
추가 배양이 끝난 후 시료가 들어있는 배양액을 제거하고 PBS용액으로 배지 성분을 제거한 후 RNAiso Plus (Takara) 용액을 1ml씩 첨가한 다음 튜브로 각 각 옮겨 클로로포름(Chloroform)용액을 200 ㎕씩 각 튜브에 첨가하고 볼텍스믹서(Vortex mixer)를 이용하여 충분히 섞어 준 후 실온에서 10분간 반응시켰다. 다음으로는 13000g, 4℃ 조건에서 15분간 원심분리하여 aqueous phase 만 새로운 튜브에서 500 ㎕ 아이소프로판올(isopropanol)과 섞어준 후 상온에서 10분간 반응시켰다. 다시 같은 조건으로 원심분리 후 상층액을 제거한 후 75% 에탄올을 첨가한 후 다시 같은 조건으로 원심분리 후 상층액인 에탄올을 완전히 제거 후 말려주었다. 각 각 튜브에 DEPC-treated Water를 40 ㎕씩 첨가 후 RNA농도를 측정하여 동일 농도로 cDNA 합성 키트(TAKARA)를 이용하여 cDNA를 합성하였다. 합성된 cDNA는 각각의 Primer 들과 함께 역전사 중합효소 연쇄반응(Reverse Transcription Polymerase Chain Reaction; RT-PCR)에 사용되어 각 각의 유전자 발현량을 1% Agarose gel에서 확인하였다.After additional incubation, remove the culture solution containing the sample, remove the medium components with PBS solution, add 1 ml of RNAiso Plus (Takara) solution, and then transfer to each tube and add 200 μl of chloroform solution to each tube. After addition, the mixture was sufficiently mixed using a vortex mixer and then reacted at room temperature for 10 minutes. Next, after centrifugation for 15 minutes at 13000 g, 4 ℃ condition, only the aqueous phase was mixed with 500 μl isopropanol (isopropanol) in a new tube, and then reacted at room temperature for 10 minutes. After centrifugation under the same conditions, the supernatant was removed, 75% ethanol was added, and then centrifuged again under the same conditions, and the ethanol as the supernatant was completely removed and dried. After adding 40 μl of DEPC-treated water to each tube, the RNA concentration was measured and cDNA was synthesized using the cDNA synthesis kit (TAKARA) at the same concentration. The synthesized cDNA was used for Reverse Transcription Polymerase Chain Reaction (RT-PCR) together with each primer, and the expression level of each gene was confirmed on 1% agarose gel.
와이셀라 용해물이 미세먼지에 의해 증가되는 MMPs의 발현에 대한 영향을 확인한 결과, 도 3에 나타낸 바와 같이, 미세먼지는 인간 각질형성세포에서 MMP-3 및 MMP-9의 유전자 발현을 증가시켰으나, 와이셀라 용해물을 전처리한 세포에서는 미세먼지에 의한 MMP-3 및 MMP-9의 증가된 발현을 억제함을 확인하였다. As a result of confirming the effect of the Wasella lysate on the expression of MMPs increased by fine dust, as shown in FIG. 3 , fine dust increased the gene expression of MMP-3 and MMP-9 in human keratinocytes, but It was confirmed that the increased expression of MMP-3 and MMP-9 caused by fine dust was inhibited in the cells pretreated with the Wasella lysate.
MMPs는 콜라겐 분해효소의 발현을 증가시켜 콜라겐을 분해함으로써 피부 주름을 유발한다. 따라서, 와이셀라 용해물이 미세먼지에 의해 감소된 콜라겐 유전자의 발현에 영향을 확인한 결과, 도 4에 나타낸 바와 같이, 미세먼지는 인간 섬유아세포 및 인간 진피 섬유아세포에서 콜라겐 유전자의 발현을 감소시켰으나, 와이셀라 용해물을 전처리한 세포에서는 미세먼지에 의한 콜라겐 유전자의 발현 감소를 억제함을 확인하였다. MMPs increase the expression of collagen-degrading enzymes to break down collagen, thereby causing skin wrinkles. Therefore, as a result of confirming the effect of the Wasella lysate on the expression of the collagen gene reduced by fine dust, fine dust reduced the expression of the collagen gene in human fibroblasts and human dermal fibroblasts, as shown in FIG. 4 , In the cells pretreated with the Wasella lysate, it was confirmed that the decrease in the expression of collagen genes caused by fine dust was suppressed.
따라서, 상기 실험 결과를 통해 와이셀라 용해물은 미세먼지에 의해 증가된 MMPs의 발현을 억제시키고, 증가된 MMPs로 인한 콜라겐을 분해를 억제하여 피부 노화 방지 및 피부 주름 개선 효과를 갖는 것을 알 수 있었다.Therefore, through the above experimental results, the Wasella lysate inhibited the expression of MMPs increased by fine dust and inhibited the decomposition of collagen caused by the increased MMPs, thereby preventing skin aging and improving skin wrinkles. .
상기와 같은 결과는 본 발명의 와이셀라 시바리아 용해물이 피부노화방지 및 피부주름 개선 효과를 갖는것을 시사하는 것이다. The above results suggest that the Wasella civaria lysate of the present invention has an effect of preventing skin aging and improving skin wrinkles.
이제까지 본 발명에 대하여 그 바람직한 실시예들을 중심으로 살펴보았다. 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들은 한정적인 관점이 아니라 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허 청구범위에 나타나 있으며, 그와 동등한 범위 내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.So far, the present invention has been looked at with respect to preferred embodiments thereof. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.
화장료의 제조예Preparation example of cosmetics
본 발명에 따른 와이셀라 시바리아 용해물은 목적에 따라 여러 형태의 화장료로 제조 가능하다. 하기는 본 발명에 따른 와이셀라 시바리아 용해물을 활성성분으로 함유시킨 몇몇 화장료의 제조방법을 예시한 것으로 본 발명이 이에 한정되는 것은 아니다.The Weissella civaria lysate according to the present invention can be prepared into various types of cosmetics depending on the purpose. The following exemplifies the manufacturing method of several cosmetics containing the Wasella civaria lysate according to the present invention as an active ingredient, but the present invention is not limited thereto.
<화장료 제조예 1> 유연 화장수의 제조<Cosmeceutical Preparation Example 1> Preparation of softened lotion
와이셀라 시바리아 용해물 10.0 중량부Weissella civaria lysate 10.0 parts by weight
1,3-부틸렌글리콜 1.00 중량부1,3-butylene glycol 1.00 parts by weight
디소듐이디티에이 0.05 중량부Disodium EDTA 0.05 parts by weight
알란토인 0.10 중량부0.10 parts by weight of allantoin
디포타슘글리시리제이트 0.05 중량부Dipotassium glycyrrhizate 0.05 parts by weight
시트르산 0.01 중량부citric acid 0.01 parts by weight
소듐시트레이트 0.02 중량부0.02 parts by weight of sodium citrate
글리세레스-26 1.00 중량부Glycereth-26 1.00 parts by weight
알부틴 2.00 중량부arbutin 2.00 parts by weight
하이드로제네이티드캐스터오일 1.00 중량부1.00 parts by weight of hydrogenated castor oil
에탄올 30.0 중량부Ethanol 30.0 parts by weight
보존제 미량preservative trace
착색제 미량trace amount of colorant
착향제 미량Trace amount of flavoring agent
정제수 잔량Purified water remaining
<화장료 제조예 2> 영양 크림의 제조<Cosmeceutical Preparation Example 2> Preparation of nourishing cream
와이셀라 시바리아 용해물 10.0 중량부Weissella civaria lysate 10.0 parts by weight
1,3-부틸렌글리콜 7.00 중량부1,3-butylene glycol 7.00 parts by weight
글리세린 1.00 중량부1.00 parts by weight of glycerin
D-판테놀 0.10 중량부D-panthenol 0.10 parts by weight
식물 추출물 3.20 중량부3.20 parts by weight of plant extract
마그네슘알루미늄실리케이트 0.30 중량부0.30 parts by weight of magnesium aluminum silicate
PEG-40 스테아레이트 1.20 중량부1.20 parts by weight of PEG-40 stearate
스테아르산 2.00 중량부2.00 parts by weight of stearic acid
폴리소르베이트 60 1.50 중량부1.50 parts by weight of
친유형글리세릴스테아레이트 2.00 중량부2.00 parts by weight of lipophilic glyceryl stearate
소르비탄세스퀴올리에이트 1.50 중량부1.50 parts by weight of sorbitan sesquioleate
세테아릴알코올 3.00 중량부3.00 parts by weight of cetearyl alcohol
미네랄오일 4.00 중량부4.00 parts by weight of mineral oil
스쿠알란 3.80 중량부Squalane 3.80 parts by weight
카르릴릭/카프릭트리글리세라이드 2.80 중량부2.80 parts by weight of carlylic/capric triglyceride
식물성오일 1.80 중량부1.80 parts by weight of vegetable oil
디메치콘 0.40 중량부Dimethicone 0.40 parts by weight
디포슘글리시리제이트 미량Trace amount of dipotassium glycyrrhizate
알란토인 미량allantoin traces
소듐 히아루로네이트 미량Sodium Hyaluronate Trace
토코페릴아세테이트 적량Tocopheryl Acetate Appropriate amount
트리에탄올아민 적량Appropriate amount of triethanolamine
보존제 적량Preservative amount
착향제 적량Appropriate amount of flavoring agent
정제수 잔량Purified water remaining
건강식품의 제조예Manufacturing example of health food
본 발명에 따른 상기 와이셀라 시바리아 용해물은 목적에 따라 여러 형태의 건강식품으로 제조 가능하다. 하기는 본 발명에 따른 와이셀라 시바리아 용해물을 활성성분으로 함유시킨 몇몇 건강식품의 제조방법을 예시한 것으로 본 발명이 이에 한정되는 것은 아니다.The Weissella civaria lysate according to the present invention can be prepared into various types of health food depending on the purpose. The following exemplifies the manufacturing method of several health foods containing the Weissella civaria lysate according to the present invention as an active ingredient, but the present invention is not limited thereto.
<건강식품 제조예 1> 유제품(dairy products)의 제조<Health food production example 1> Production of dairy products
본 발명의 와이셀라 시바리아 용해물 0.01-1 중량부를 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.0.01-1 parts by weight of the Weissella civaria lysate of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.
<건강식품 제조예 2> 선식의 제조<Health food production example 2> Preparation of wire food
현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화시켜 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다. 검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다. 본 발명의 와이셀라 시바리아 용해물을 진공 농축기에서 감압농축하고 건조분말을 얻었다. 상기에서 제조한 곡물류, 종실류 및 와이셀라 시바리아 용해물의 건조분말을 다음의 비율로 배합하여 제조하였다.Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared into powder having a particle size of 60 mesh with a grinder. The Wasella civaria lysate of the present invention was concentrated under reduced pressure in a vacuum concentrator to obtain a dry powder. It was prepared by mixing the dry powder of the grains, seeds and Weissella civaria lysate prepared above in the following ratio.
곡물류(현미 34 중량부, 율무 19 중량부, 보리 20 중량부),Grains (34 parts by weight of brown rice, 19 parts by weight of barley, 20 parts by weight of barley),
종실류(들깨 7 중량부, 검정콩 8 중량부, 검정깨 7 중량부),Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame),
와이셀라 시바리아 용해물 (2 중량부),Weissella civaria lysate (2 parts by weight),
영지(1.5 중량부), 및Reishi (1.5 parts by weight), and
지황(1.5 중량부).Rehmannia (1.5 parts by weight).
건강기능식품의 제조예Manufacturing example of health functional food
본 발명에 따른 와이셀라 시바리아 용해물은 목적에 따라 여러 형태의 건강기능식품으로 제조 가능하다. 하기는 본 발명에 따른 와이셀라 시바리아 용해물을 활성성분으로 함유시킨 몇몇 건강기능식품의 제조방법을 예시한 것으로 본 발명이 이에 한정되는 것은 아니다.The Weissella civaria lysate according to the present invention can be prepared into various types of health functional food depending on the purpose. The following exemplifies the manufacturing method of several health functional foods containing the Wasella civaria lysate according to the present invention as an active ingredient, but the present invention is not limited thereto.
<건강기능식품 제조예 1> 건강기능식품의 제조<Health functional food manufacturing example 1> Manufacturing of health functional food
와이셀라 시바리아 용해물 100 mgWeissella civaria lysate 100 mg
비타민 혼합물 적량appropriate amount of vitamin mixture
비타민 A 아세테이트 70 μgVitamin A Acetate 70 μg
비타민 E 1.0 mgVitamin E 1.0 mg
비타민 B1 0.13 mgVitamin B1 0.13 mg
비타민 B2 0.15 mgVitamin B2 0.15 mg
비타민 B6 0.5 mg0.5 mg of vitamin B6
비타민 B12 0.2 μg0.2 μg of vitamin B12
비타민 C 10 mg
비오틴 10 μgBiotin 10 μg
니코틴산아미드 1.7 mgNicotinamide 1.7 mg
엽산 50 μg50 μg of folic acid
판토텐산 칼슘 0.5 mgCalcium pantothenate 0.5 mg
무기질 혼합물 적량Mineral mixture appropriate amount
황산제1철 1.75 mgferrous sulfate 1.75 mg
산화아연 0.82 mgZinc Oxide 0.82 mg
탄산마그네슘 25.3 mgMagnesium carbonate 25.3 mg
제1인산칼륨 15 mgpotassium phosphate monobasic 15 mg
제2인산칼슘 55 mgDibasic calcium phosphate 55 mg
구연산칼륨 90 mgPotassium citrate 90 mg
탄산칼슘 100 mg100 mg of calcium carbonate
염화마그네슘 24.8 mgMagnesium chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강기능성 식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강기능성 식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강기능성 식품 조성물 제조에 사용할 수 있다.Although the composition ratio of the vitamin and mineral mixture is relatively suitable for health functional food in a preferred embodiment, the composition ratio may be arbitrarily modified. Then, the granules can be prepared and used in the production of a health functional food composition according to a conventional method.
<건강기능식품 제조예 2> 건강 기능 음료의 제조<Health functional food production example 2> Preparation of health functional beverage
와이셀라 시바리아 용해물 100 mgWeissella civaria lysate 100 mg
구연산 100 mg100 mg citric acid
올리고당 100 mg100 mg of oligosaccharides
매실농축액 2 mg
타우린 100 mg
정제수를 가하여 전체 500 mLAdd purified water to total 500 mL
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 1 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다. 상기 조성비는 비교적 기호 음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.After mixing the above ingredients according to a conventional health drink manufacturing method, after stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized container, sealed and sterilized, and then refrigerated. used in the manufacture of health beverage compositions of Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demand country, and use purpose.
Claims (11)
상기 피부 주름은 미세먼지에 의한 것을 특징으로 하는 피부 주름 개선 또는 탄력증진용 화장료 조성물.The method of claim 1,
The skin wrinkles are a cosmetic composition for improving skin wrinkles or enhancing elasticity, characterized in that due to fine dust.
상기 화장료 조성물은 스킨로션, 스킨소프너, 스킨토너, 아스트린젠트, 로션, 밀크로션, 모이스쳐 로션, 영양로션, 맛사지크림, 영양크림, 모이스처크림, 핸드크림, 파운데이션, 에센스, 영양에센스, 팩, 비누, 클렌징폼, 클렌징로션, 클렌징크림, 바디로션 및 바디클린저로 구성된 제형에서 선택된 하나 이상의 제형인 것을 특징으로 하는 피부 주름 개선 또는 탄력증진용 화장료 조성물.The method of claim 1,
The cosmetic composition includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, soap, cleansing A cosmetic composition for improving skin wrinkles or enhancing elasticity, characterized in that it is one or more formulations selected from formulations consisting of foam, cleansing lotion, cleansing cream, body lotion and body cleanser.
상기 건강기능식품은 정제, 캡슐제, 환제 또는 액제 형태의 식품인 것을 특징으로 하는 피부 주름 개선 또는 탄력증진용 건강기능식품 조성물.7. The method of claim 6,
The health functional food is a health functional food composition for improving skin wrinkles or enhancing elasticity, characterized in that the food is in the form of tablets, capsules, pills or liquids.
상기 건강식품은 각종 드링크제, 육류, 소세지, 빵, 캔디류, 스넥류, 면류, 아이스크림, 유제품, 스프, 이온음료, 음료수, 알코올 음료, 껌, 차 및 비타민 복합제에서 선택되는 것을 특징으로 하는 피부 주름 개선 또는 탄력증진용 건강식품 조성물.10. The method of claim 9,
The health food is selected from various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic beverages, beverages, alcoholic beverages, gum, tea and vitamin complexes. Skin wrinkle improvement or A health food composition for improving elasticity.
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